Biochemical Analysis of the Arginine Methylation of High Molecular Weight Fibroblast Growth Factor-2

• Published in: The Journal of biological chemistry Vol. 275; no. 5; pp. 3150 - 3157
• Main Authors: Klein, Sharon, Carroll, James A., Chen, Yan, Henry, Michael F., Henry, Pamela A., Ortonowski, Izabela E., Pintucci, Giuseppe, Beavis, Ronald C., Burgess, Wilson H., Rifkin, Daniel B.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 04.02.2000; American Society for Biochemistry and Molecular Biology
• Subjects: 3T3 Cells; Amino Acid Sequence; Animals; Arginine - chemistry; Fibroblast Growth Factor 2 - chemistry; Fibroblast Growth Factor 2 - genetics; Fibroblast Growth Factor 2 - metabolism; Humans; Mass Spectrometry; Methylation; Mice; Molecular Sequence Data; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Sequence Analysis, Protein
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.275.5.3150

The post-translational methylation of the N-terminally extended or high molecular weight (HMW) forms of fibroblast growth factor-2 (FGF-2) has been shown to affect the nuclear accumulation of the growth factor. In this study, we determined the extent and position of methyl groups in HMW FGF-2. Using mass spectrometry and amino acid sequence analysis, we have shown that the 22- and 22.5-kDa forms of HMW FGF-2 contain five dimethylated arginines located at positions −22, −24, −26, −36, and −38 using the methionine residue normally used to initiate the 18-kDa form as position 0. The 24-kDa form of HMW FGF-2 contains seven to eight dimethylated arginines located at positions −48, −50, and −52, in addition to positions −22, −24, −26, −36, and −38.In vitro methylation reactions demonstrate that the N-terminal extension of HMW FGF-2 acts as a specific substrate for yeast Hmt1p and human HRMT1L2 arginine methyltransferases. These findings indicate that HMW FGF-2, with the presence of five or more dimethylated Gly-Arg-Gly repeats, contains an RGG box-like domain, which may be important for protein-protein and/or protein-RNA interactions.